Alterations in the tegument surface of adult fluke, Fasciola gigantica, after in vitro treatment with the crude venom of the oriental hornet, Vespa orientalis

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Alterations in the tegument surface of adult fluke, Fasciola gigantica, after in vitro treatment with the crude venom of the oriental hornet, Vespa orientalis

https://doi.org/10.21203/rs.3.rs-2116577/v1

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Triclabendazole is the only known effective treatment against both pre-adult and adult worms infecting the hepatic parenchyma and bile ducts, respectively. We aimed to investigate the antiparasitic activity of the venom of the wasp, Vespa orientalis (WV), against the adult Fasciola gigantica (F. gigantica) worm, as WV has known pharmacological properties. We assessed its anthelmintic efficacy using 40 adult F. gigantica flukes collected from slaughtered cattle divided into four groups (10 each). Three worms in each group were treated with 50, 100, and 200 µg/ml of WV, while a fourth untreated one was used as control. After WV treatment, the tegument area of the fluke’s body was assessed using a scanning electron microscope (SEM). This revealed several tegumental alterations all over the fluke's body, including sunken spines due to local swelling, complete disappearance of spines from their sockets, furrowing and sloughing of the basal lamina, splitting off some spines to resemble an “open jaw,” broken oral and ventral sucker teguments, increased swelling of the ventral sucker with loss of its ridges, severe swelling and smoothening of the ventral sucker due to loss of normal transverse ridges, deformed cirrus, and swollen sensory papillae with blebs. The sensory papillae were completely disrupted and dislodged, leaving pits and a series of holes throughout the basal lamina and severely eroded, turning into lesions exposing the basement membrane. These findings indicate that WV can be potentially used to treat fascioliasis. However, further in vivo studies on the activity of WV treatment are recommended.

Tegument surface alteration

Fasciola gigantica

Vespa orientalis

venom

The venom of the oriental hornet, Vespa orientalis, has several physical and pharmacological properties as it contains bioactive compounds, including small peptides, amines, and low/high molecular weight proteins (enzymes and toxins) (Nakajima 1986). The biological effects of this venom can be attributed to the presence of serotonin, acetylcholine, kinins, adrenaline, noradrenaline, dopamine, and high molecular weight compounds such as hyaluronidase, phospholipase-A2, histidine decarboxylase, poly-, and disaccharidases, neutral DNAse, and several polycationic peptides (Piek 1986; Abd El-Wahed et al. 2021).

Fascioliasis is an important helminthic disease caused by the parasitic liver fluke species from the genus Fasciola (Digenea: Fasciolidae) (Waikagul et al. 2015). F. gigantica and F. hepatica significantly impact the growth rate, development, and productivity of ruminants and thus are economically important (Beesley et al. 2018). Fascioliasis is treated exclusively using various anthelminthic drugs (Ullah et al. 2017). Since 1983, the benzamide compound “Triclabendazole” has been the only approved drug for veterinary and human use. It was recommended by the WHO in 1997, especially for endemic countries with a high fascioliasis burden (McCarthy and Moore 2015). Triclabendazole is the only known drug effective against the pre-adult and adult worms of F. gigantica in the hepatic parenchyma and bile ducts, respectively. However, several studies have reported resistance to this drug (Cabada et al. 2016; Gandhi et al. 2019). Currently, novel, effective alternative treatments for fascioliasis are urgently needed due to the increased prevalence of drug resistance. Natural alternative therapies are being explored due to their higher biosafety and fewer side effects than synthetic drugs (Ullah et al. 2017).

In this study, we used scanning electron microscopy (SEM) to investigate the in vitro effect of the wasp venom on the adult Fasciola gigantica worms and to assess its therapeutic potential.

2.1. Collection and preparation of parasites

The adult liver flukes, F. gigantica, were either collected from the bile ducts or the gallbladders of the animals slaughtered at the Drunka altar in the Assiut governorate, Egypt. They were washed several times with 0.85% NaCl solution, and only intact and actively mobile flukes were selected for the study.

2.2. Collection of wasps’ crude venom

The wasps were collected by placing traps among the honeybee hives at the fields of the Faculty of Agriculture, Al-Azhar University in Assiut, from October to December 2021. To obtain the venom, the wasps were immobilized by rapid freezing at −20°C. Then, the stinging apparatus were dissected at 4°C, and the venom reservoirs were extracted and stored at −20°C until further use. The venom sacs were resuspended in deionized water (MilliQ). The whole wasp venom extract was obtained by disrupting the reservoirs using a glass rod under rapid defrosting and light pressure. The samples were centrifuged at 10,000 g for 5 minutes at 4°C, and the supernatants were used as protein and enzyme sources after lyophilization. The lyophilized crude wasp venom was dissolved in MilliQ water and filtered through a 0.22 µm syringe filter (Surendra et al. 2011). The worms were incubated in vitro in sterile RPMI-1640 medium (Caisson Laboratories Inc. Smithfield, UT, USA) supplemented with L-glutamine, penicillin (50 IU/ml), streptomycin (50 µg/ml), and gentamycin (30 IU/ml) (Sigma-Aldrich, St. Louis, Missouri, USA). The working solutions of WV (50, 100, and 200 µg/ml) were prepared by dissolving it in RPMI-1640 complete medium as previously described (Nair et al. 2016).

2.3. Study design

The F. gigantica adults were kept under sterile conditions inside a laminar flow cabinet in a petri dish containing RPMI-1640 complete medium. The worms were divided into four groups (10 flukes/group) as follows; Group 1 (control), the worms were incubated in the medium without WV; Group 2, 3, and 4 were incubated in the medium with 50, 100, and 200 µg/ml of WV, respectively for 24 hours at 37°C (Sumsakul et al. 2014).

2.4. SEM

The F. gigantica adult liver flukes were examined using SEM as previously described (Shalaby et al. 2016). Briefly, the samples were fixed in 2.5% buffered glutaraldehyde in 0.1 M PBS (pH 7.4) at 4°C for 24 hours, washed with PBS, and re-fixed with 1% osmium tetroxide for one hour. The samples were rewashed with PBS, dehydrated using a serial dilution of ethyl alcohol (30, 50, 70, 90, and 100%), and infiltrated with acetone. The samples were dried with SPI Critical Point Dryer and mounted on gold-coated aluminum stubs in an SPI Module ™ Vac/Sputter coater (SPI Supplies, West Chester, PA, USA). Finally, the specimens were examined by the JSM-52500 LV SEM (JEOL Ltd. Inc., Tokyo, Japan) at the Electron Microscope Unit of Assiut University.

3.1. SEM of the control flukes

The morphology of the control group (Figs. 1a, 2a, 3a, 4a, 4b, 5a, and 6a) showed leaf-like bodies with tapered anterior and posterior ends. The apical cone was located anteriorly with an oral sucker at its tip. A ventral sucker was present on the ventral surface at the apical cone-main body junction with the gonopore just anterior to it (Fig. 1a). The typical intact tegument surface exhibited micro ridges and grooves with numerous serrated spines, which were deficient at the rims of both suckers (Fig. 2a). A cluster of fungiform-shaped sensory papillae surrounded the ventral sucker. Also, these papillae were scattered throughout the body’s surface between the spines (Fig. 3a and 4b). Each papilla has a small dome with a smooth top and a highly pitted base. The spines were mainly located on the anterior part, especially around the oral and ventral suckers, and were fewer on the posterior part of the body. The size and shape of these spines depended on their location on the fluke’s body. The spines on the apical cone were broad, tightly packed, and posteriorly directed, with several fingers-like protrusions at their tips (Figs. 4a and b). The spines on the mid-body were not as closely arranged as those on the apical cone, and the protrusions along their tips were not well-defined (Fig.5a). The spines on the tail surface were smaller than the rest (Fig. 6a).

3.2. The effects of WV on adult Fasciola gigantica

3.2.1. After incubation with 50 µg/ml WV for 24 hr

After 24 hr incubation with 50 µg/ml WV, the tegumental surface of the apical cone was severely swollen with exposed basal lamina and deep folds (Fig. 1b). The ventral sucker showed increased swelling and loss of ridges (Fig. 2b). The sensory papillae were swollen and covered with blebs, including a few disrupted ones (Fig. 3b). The tegument was sloughed, and few spines were split, appearing like “open jaws” (Fig. 4c). The mid-body region showed tegumental swelling in the interspinal region with the split, “open-jawed” spines (Fig. 5c). The spines in the tail region often appeared sunken due to the swelling in the surrounding tegument (Fig. 6b).

3.2.2. After incubation with 100 µg/ml WV for 24 h

After 24 h incubation with 100 µg/ml WV, the oral suckers in the apical cone region showed damaged teguments with deformed cirrus. The tegumental surface was severely swollen with exposed basal lamina (Fig. 1c). The ventral sucker was severely swollen and appeared smooth due to the loss of the normal transverse ridges around its rim and sloughing of the tegument, exposing basal lamina underneath (Fig. 2c). All the sensory papillae were disrupted and dislodged from their places, leaving pits (Fig. 3c). The spines were also completely damaged and dislodged, leaving behind empty spine sockets (Fig. 4d). The mid-body region showed submerged spines, which were either completely flattened against the surface or submerged in the swollen tegument, leaving deep furrows around them (Fig. 5c and d). In the tail region, the spines disappeared, leaving empty spine sockets (Fig. 6c).

3.2.3. After incubation with 200 µg/ml WV for 24 h

After 24 h incubation with 200 µg/ml WV, the apical cone region showed damaged tegument of the oral and ventral suckers, deformed cirrus, severely swollen tegumental surface with exposed basal lamina and furrows (Fig. 1d and Fig. 2d). A series of holes were observed around the basal lamina with the sloughed-off surrounding tegument and complete disappearance of the sensory papillae clusters (Fig. 3d). The tegument of the anterior region was severely eroded, eventually turning into lesions, exposing a large area of the basement membrane and complete loss of spines (Fig. 4e). The mid-body region displayed furrowed tegument, complete loss of spines and sloughed tegument with exposed underlying basal lamina (Fig. 5e). The tail region was also furrowed with complete disappearance of spines, leaving spine sockets and sloughed tegument, exposing the basal lamina (Fig. 6d).

Fascioliasis treatment is dependent on the use of chemical drugs (Saowakon et al. 2009). However, excessive and continuous usage of these drugs has given rise to drug resistance, which has been reported in many endemic regions globally (Alvarez-Mercado et al. 2015; McCarthy and Moore 2015). Currently, medical and pharmacological research is focused on investigating novel, natural treatments because of their higher biosafety levels and lower side effects than synthetic drugs (Alvarez-Mercado et al. 2015; Toner et al. 2010). Animal venoms are abundant in natural bioactive molecules with excellent pharmaceutical and therapeutic properties. Thus they can be potentially used for drug discovery (Rivero et al. 2011; Vigerelli et al. 2014; Neto et al. 2015). Hassan et al. (2016) studied the potential anti-schistosomal activity of the Egyptian snake, Cerastes cerastes venom, in vitro. They found that it causes mild to severe tegumental damage to Schistosoma worms. Al-Malki and Abdelsater (2020) reported that 100 µg/mL of crude venom from the scorpion, Androctonus crassicauda (ACCV), can destroy all Echinococcus granulosus protoscolices after 240 min incubation. Al-Malki et al. (2022) also reported that ACCV induces severe ultrastructural changes and apoptosis of Echinococcus granulosus protoscolices. Kellershohn et al. (2019) assessed the antischistosomal capacity of harmonine, an antimicrobial alkaloid from the harlequin ladybird Harmonia axyridis. They observed that it had remarkable pleiotropic effects on the physiological, cellular, and molecular processes in adult male and female Schistosoma mansoni. Tonk et al. (2020) investigated the antischistosomal activity of the venom from the European predatory assassin bug, Rhynocoris iracundus. The components of wasp venom are commonly categorized as (i) bioactive molecules such as serotonin, histamine, catecholamines, tyramine, acetylcholine, flavonoids, and biologically active amines, (ii) high molecular weight proteins such as hyaluronidases, phospholipases, and antigen 5, and (iii) low molecular weight peptides such as mastoparan (Nakajima 1986). Mastoparan, a wasp kinin and a chemotactic peptide is a known tetra decapeptide present in wasp venom (Hoffman and Jacobson 1984) that can create pores in the lipid bilayers of erythrocytes, bacteria, and mast cells (Hoffman and Jacobson, 1984). Moreover, it can also induce pore formation and disruption of the viral lipid envelope (Sample et al. 2013). Vespoid wasp phospholipases can disrupt the phospholipid layer of several biological membranes, leading to pore formation and/or cell lysis (Santos et al. 2007; Costa and Palma 2000). The development of electron microscopy has facilitated the morphological analysis of parasites, which helps understand the complex morphological changes due to the parasite and drug-parasite interactions (Sachanonta et al. 2011). SEM is used to evaluate the efficacy of many fasciolicides (Anderson and Fairweather 1998; Buchanan et al. 2002; Meaney et al. 2002, 2003) because it enables observation of the damaged surface of the fluke tegument, which has different mechanical and metabolic functions in the parasite (Dalton 1999). To our knowledge, this study is the first to investigate the effect of wasp venom on the tegument surface of adult F. gigantica in a dose-dependent manner. Our SEM data demonstrated various tegumental, including tegument swelling, the disappearance of spines, damaged tegument of the oral and ventral suckers, deformed cirrus, destroyed and displaced sensory papillae from their places, a series of holes in the basal lamina, splitting of spines (opened jaw), severe erosion, the complete loss of spines, furrowing, folding, sloughing, and blebbing of the tegument. The severity increased with an increase in doses of wasp venom. The tegumental damage might induce extensive alterations in the underlying tissues and internal organs, causing more damage to the entire parasite (Shalaby et al. 2016). The wasp venom (WV) also disrupts several physiological processes associated with the tegument, including osmoregulation, nutrient uptake, secretion, synthesis, and immune protection (Massoud et al. 2013). The destruction of the spines and sensory papillae will undoubtedly deprive the parasite of its vital functions and facilitate its clearance. Consistent with our findings, several studies used various plant and animal sources to reveal the same effects. For instance, Keiser and Morson (2008) and Shalaby et al. (2009) studied the effects of some plant extracts on the adult Fasciola tegument ultrastructure. Saowakon et al. (2009) studied the impact of Artocarpus lakoocha aqueous extract on the tegument of adult F. gigantica. Shehab et al. (2009) studied the in vitro fasciolicidal activity of Meryta denhamii plant extracts against adult Fasciola. Ebeid et al. (2011) determined the in vitro effects of plant extracts from B. egyptiaca compared with TCBZ in adult F. gigantica. Shoheiba and Darb (2011) evaluated both tegument alterations in adult F. gigantica and egg hatchability after in vitro incubation with either Commiphora molmol (C. molmol) or Nigella sativa (N. sativa) extract. Ullah et al. (2017) studied the anthelmintic and fasciolicidal activity of both curcumin and thymoquinone on the tegument of the adult F. gigantica. Lorsuwannarat et al. (2014) and Abdel Fattah et al. (2021) suggested that a Plumbagin compound has a fasciolicidal activity on F. gigantica.

Excessive and continual usage of chemical drugs has resulted in drug resistance, which has been reported in many endemic regions of the world. Therefore, medical and pharmacological research focuses on investigating novel therapeutics from natural sources as they are safer and have fewer side effects than chemicals. Animal venoms are important sources of natural bioactive molecules with significant therapeutic potential. Our findings revealed that the Vespa orientalis venom (WV) has a promising fasciolicidal activity as it can significantly alter the teguments of the adult F. gigantica. These results facilitate the development of novel approaches toward controlling and treating fascioliasis using novel trematocidal agents. However, further studies need to be conducted to characterize and identify this venom and its active compound (s) responsible for the fasciolicidal activity and to understand its mechanism of action.

Competing interests: The authors declare that there is no conflict of interest.

Acknowledgments

The authors would like to thank Dr. Mostafa Abozeid, the veterinarian at Assiut abattoir, for his assistance in collecting the adult Fasciola samples from the infected animals.

Abdel Fattah D, Temsah KA, El Kholy WAS, El Kholy AA, El-Dardiry MA (2021). Fasciola gigantica: tegumental surface alterations following treatment in vitro with Plumbagin. Al-Azhar Un. J Res Stud 3.

Abd El-Wahed A, Yosri N, Sakr HH, Du M, Algethami AFM, Zhao C, Abdelazeem AH, Tahir HE, Masry SHD, Abdel-Daim MM, Musharraf SG, El-Garawani I, Kai G, Khalifa SAM, El-Seedi HR (2021) Wasp venom biochemical components and their potential in biological applications and nanotechnological interventions. Toxins 13:206.

Al-Malki ES, Abdelsater N (2020) In vitro scolicidal effects of Androctonus crassicauda (Olivier, 1807) venom against the protoscolices of Echinococcus granulosus. Saudi J Biol Sci 27:1760-1765.

Al-Malki ES, Aljedaie MM, Amer OSO, Abdelsater N, Badry A (2022) Scorpion crude venom induced apoptosis and structural changes of Echinococcus granulosusprotoscolices. J King Saud Univ Sci 34:101937. https://doi.org/10.1016/jjksus.2022.101937.

Alvarez-Mercado JM, Ibarra-Velarde F, Alonso-Diaz MA, Vera-Montenegro Y, Avila-Acevedo JG, Garcia-Bores AM (2015) In vitro antihelmintic effect of fifteen tropical plant extracts on excited flukes of Fasciola hepatica. BMC Vet Res 11:45.

Anderson HR, Fairweather I (1998) Fasciola hepatica: scanning electron microscopic evaluation of juvenile flukes following treatment in vitro with deacetyllated (amine) metabolite of diamphenethide(damd). Int J Parasitol 18:827-837.

Beesley NJ, Caminade C, Charlier J et al (2018) Fasciola and fasciolosis in ruminants in Europe: identifying research needs. Transbound Emerg Dis 65:199-216.

Buchanan JF, Fairweather I, Brennan GP, Trudget A, Hoey EM (2002) Fasciola hepatica: surface and tegumental changes inducedby treatment in vitro with the sulphoxide metabolite of albendazole (valbazen). Parasitology 126:141-153

Cabada MM, Lopez M, Cruz M, Delgado JR, Hill V, White AC Jr (2016) Treatment failure after multiple courses of triclabendazole among patients with Fascioliasis in Cusco, Peru: a case series. PLOS Negl Trop Dis 10:e0004361.

Costa H, Palma MS (2000) Agelotoxin: a phospholipase A2 from the venom of the neotropical social wasp cassununga (Agelaiapallipespallipes) (Hymenoptera-Vespidae). Toxicon 38:1367-1379.

Dalton JP (1999) Fasciolosis. University Press, New York

Ebeid MH, Moustafa AM, Arnaout FK et al (2011) In vitro evaluation of anthelmentic efficacy of balanitesegyptiaca on fasciolagigantica. BVMJ 22:56-67.

Gandhi P, Schmitt EK, Chen CW, Samantray S, Venishetty VK, Hughes D (2019) Triclabendazole in the treatment of human fascioliasis: a review. T Roy Soc Trop Med H 113:797-804.

Hassan EA, Abdel-Rahman MA, Ibrahim MM, Soliman MFM (2016) In vitro antischistosomal activity of venom from the Egyptian snake Cerastes cerastes. Rev Soc Bras Med Trop 49:752-757. https://doi.org/10.1590/0037-8682-0241-2016.

Hoffman DR, Jacobson RS (1984) Allergens in hymenoptera venom XII: how much protein is in a sting? Ann Allergy 52:276-278.

Keiser J, Morson G (2008). Fasciola hepatica: surface tegumental responses to in vitro and in vivo treatment with the experimental fasciolicide OZ 78. Exp Parasitol 119:87- 93.

Kellershohn J, Thomas L, Gru’nweller A, Hartmann RK, Hardt M, Vilcinskas A et al (2019) Insects in anthelminthics research: lady beetlederivedharmonine affects survival, reproduction and stem cell proliferation of Schistosomamansoni. PLoS Negl Trop Dis 13: e0007240. https://doi.org/10.1371/journal.pntd.0007240

Lorsuwannarat N, Piedrafita D, Chantree P et al (2014). The in vitro anthelmintic effects of plumbagin on newly excysted and 4-weeks- old juvenile parasites of Fasciola gigantica. Exp Parasitol 136:5-13.

Massoud AM, Shalaby HA, El Khateeb RM, Mahmoud MS, Kutkat MA (2013) Tegumental histological effects of Mirazid(®) and myrrh volatile oil on adult Fasciola gigantica. Asian Pac J Trop Biomed 3:501-504.

McCarthy JS, Moore TA (2015) 42 - Drugs for Helminths, Editor(s): John E. Bennett, Raphael Dolin, Martin J. Blaser. In: Mandell D and Bennett’s principles and practice of infectious diseases, 8th edn. Content Repository Only!, pp 519- 527.e3.

Meaney M, Fairweather I, Brennan GP, Ramasamy P, Subramanian PB (2002) Fasciola gigantica: tegumental surface alterationsfollowing treatment in vitro with the sulphoxidemetabolite of triclabendazole. Parasitol Res 88:315-325

Meaney M, Fairweather I, Brennan GP, McDowell LSL, Forbes AB (2003) Fasciola hepatica: effects of the fasciolicideclorsulonin vitro and in vivo on the tegumental surface, and a comparison of the effects on young and old mature flukes. Parasitol Res 91:238-250

Nair SV, Baranwal G, Chatterjee M et al (2016). Antimicrobial activity of plumbagin, a naturally occurring naphthoquinone from Plumbago rosea, against Staphylococcus aureus and Candida albicans. Int J Med Microbiol Suppl 306:237-248.

Nakajima T (1986) Pharmacological biochemistry of vespid venoms. In: Venoms of the Hymenoptera: Biochemical, Pharmacological and Behavioural Aspects; Piek, T., Ed.; Academic Press: London, UK, pp 309-27.

Neto RSC, Vigerelli H, Jared C, Antoniazzi MM, Chaves LB, Cassia Rodrigues da Silva A, Melo RL, Sciani JM, Pimenta DC (2015) Synergic effects between ocellatin-F1 and bufotenine on the inhibition of BHK-21 cellular infection by the rabies virus. J Venom Anim Toxins Incl Trop Dis 21:50. https://doi.org/10.1186/s40409-015-0048-1.

Piek TSW (1986) Chemistry and pharmacology of solitary wasp venoms. In: Venoms of the Hymenoptera: Biochemical, Pharmacological and Behavioural Aspects; Piek, T., Ed.; Academic Press: London, UK, pp 61-307.

Rivero JVR, de Castro FOF, Stival AS, Magalhaes MR, Carmo Filho JR, Pfrimer IAH (2011) Mechanisms of virus resistance and antiviral activity of snake venoms. J Venom Anim Toxins incl Trop Dis 17:387-393. https://doi.org/10.1590/S1678-91992011000400005.

Sachanonta N, Chotivanich K, Chaisri U et al (2011) Ultrastructural and real-time microscopic changes in P. falciparum-infected red blood cells following treatment with antimalarial drugs. Ultrastruct Pathol 35:214-225.

Sample CJ, Hudak KE, Barefoot BE, Koci MD, Wanyonyi MS, Abraham S, Staats HF, Ramsburg EA (2013) A mastoparan-derived peptide has broadspectrum antiviral activity against enveloped viruses. Peptides 48:96-105.

Santos LD, Santos KS, Souza BM, Arcuri HA, Cunha-Neto E, Castro FM, Kalil JE, Palma MS (2007) Purification, sequencing and structural characterization of the phospholipase A1 from the venom of the social wasp Polybiapaulista (Hymenoptera, Vespidae). Toxicon 1:923-937.

Saowakon N, Tansatit T, Wanichanon C, Chanakul W, Reutrakul V, Sobhon P (2009) Fasciola gigantica: anthelmintic effect of the aqueous extract of Artocarpus lakoocha. Exp Parasitol 122:289-298.

Shalaby HA, El Namaky AH, Kamel ROA (2009). In vitro effect of artemether and triclabendazole on adult Fasciola gigantica. Vet Parasitol 160:76-82

Shalaby HA, El Namaky AH, Kamel RO (2016) In vitro tegumental alterations on adult Fasciola gigantica caused by mefloquine. J Parasit Dis 40:145-151.

Shehab NG, Monem AR, Hassan SE, Toaleb NI (2009) Botanical study of Merytadenhamii Seem and its antihelminthic activity against Fasciola gigantica. J Egypt Soc Parasitol 39:269-288.

Shoheiba Z, Darb Y (2011) In vitro effect of some plant extracts on Fasciola Gigantica tegument ultrastructure and egg hatchability. Egypt J Schistosomiasis Infect Endem Dis. 33:45-58.

Sumsakul W, Plengsuriyakarn T, Chaijaroenkul W, Viyanant V, Karbwang J, Na-Bangchang K (2014) Antimalarial activity of plumbagin in vitro and in animal models. BMC Complem Altern M 14:15.

Saowakon N, Tansatit T, Wanichanon C, Chanakul W, Reutrakul V, Sobhon P (2009) Fasciola gigantica: anthelmintic effect of the aqueous extract of Artocarpus lakoocha. Exp Parasitol 122:289-298.

Surendra NS, Jayaram GN, Reddy MS (2011) Antimicrobial activity of crude venom extracts in honeybees (Apiscerana, Apisdorsata, Apisflorea) tested against selected pathogens. Afr J Microbiol Res 5:2765-2772.

Toner E, Brennan GP, Hanna RE, Edgar HW (2010) Time-dependent changes to the tegumental system and gastrodermis of adult Fasciola hepatica following treatment in vivo with triclabendazole in the sheep host. Vet Parasitol 174:218-227.

Tonk M, Vilcinskas A, Grevelding CG, Haeberlein.S (2020) Anthelminthic activity of assassin bug venomagainst the blood fluke Schistosoma mansoni. Antibiotics 9:664. https://doi.org/10.3390/antibiotics9100664

Ullah R, Rehman A, Zafeer MF et al (2017) Anthelmintic potential of thymoquinone and curcumin on Fasciola gigantica. PloS One 12:e0171267.

Vigerelli H, Sciani JM, Jared C, Antoniazzi MM, Caporale GMM, Silva ACR, Pimenta DC (2014) Bufotenine is able to block rabies virus infection in BHK-21 cells. J Venom Anim Toxins Incl Trop Dis 20:45. https://doi.org/10.1186/1678-9199-20-45.

Waikagul J, Sato M, Sato MO (2015) Foodborne trematodes (Chapter 10), Editor: Gajadhar A. A. In: Woodhead Publishing Series in Food Science, Technology and Nutrition, Foodborne Parasites in the Food Supply Web. 1st edn. Woodhead Publishing, pp 221-255.

No competing interests reported.

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Alterations in the tegument surface of adult fluke, Fasciola gigantica, after in vitro treatment with the crude venom of the oriental hornet, Vespa orientalis

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1. Introduction

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